Abstract
Misfolded proteins are associated with several pathological conditions including neurodegeneration. Although some of these abnormally folded proteins result from mutations in genes encoding disease-associated proteins (for example, repeat-expansion diseases), more general mechanisms that lead to misfolded proteins in neurons remain largely unknown. Here we demonstrate that low levels of mischarged transfer RNAs (tRNAs) can lead to an intracellular accumulation of misfolded proteins in neurons. These accumulations are accompanied by upregulation of cytoplasmic protein chaperones and by induction of the unfolded protein response. We report that the mouse sticky mutation, which causes cerebellar Purkinje cell loss and ataxia, is a missense mutation in the editing domain of the alanyl-tRNA synthetase gene that compromises the proofreading activity of this enzyme during aminoacylation of tRNAs. These findings demonstrate that disruption of translational fidelity in terminally differentiated neurons leads to the accumulation of misfolded proteins and cell death, and provide a novel mechanism underlying neurodegeneration.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Loftfield, R. B. & Vanderjagt, D. The frequency of errors in protein biosynthesis. Biochem. J. 128, 1353–1356 (1972)
Jakubowski, H. & Goldman, E. Editing of errors in selection of amino acids for protein synthesis. Microbiol. Rev. 56, 412–429 (1992)
Bacher, J. M., de Crecy-Lagard, V. & Schimmel, P. R. Inhibited cell growth and protein functional changes from an editing-defective tRNA synthetase. Proc. Natl Acad. Sci. USA 102, 1697–1701 (2005)
Doring, V. et al. Enlarging the amino acid set of Escherichia coli by infiltration of the valine coding pathway. Science 292, 501–504 (2001)
Nangle, L. A., de Crecy-Lagard, V., Doring, V. & Schimmel, P. Genetic code ambiguity. Cell viability related to the severity of editing defects in mutant tRNA synthetases. J. Biol. Chem. 277, 45729–45733 (2002)
Pezo, V. et al. Artificially ambiguous genetic code confers growth yield advantage. Proc. Natl Acad. Sci. USA 101, 8593–8597 (2004)
Selkoe, D. J. Folding proteins in fatal ways. Nature 426, 900–904 (2003)
Dobson, C. M. Protein folding and misfolding. Nature 426, 884–890 (2003)
Ross, C. A. & Poirier, M. A. Protein aggregation and neurodegenerative disease. Nature Med. 10 (Suppl.), S10–S17 (2004)
Dock-Bregeon, A. et al. Transfer RNA-mediated editing in threonyl-tRNA synthetase. The class II solution to the double discrimination problem. Cell 103, 877–884 (2000)
Beebe, K., Ribas De Pouplana, L. & Schimmel, P. Elucidation of tRNA-dependent editing by a class II tRNA synthetase and significance for cell viability. EMBO J. 22, 668–675 (2003)
Hou, Y. M. & Schimmel, P. Evidence that a major determinant for the identity of a transfer RNA is conserved in evolution. Biochemistry 28, 6800–6804 (1989)
Ripmaster, T. L., Shiba, K. & Schimmel, P. Wide cross-species aminoacyl-tRNA synthetase replacement in vivo: yeast cytoplasmic alanine enzyme replaced by human polymyositis serum antigen. Proc. Natl Acad. Sci. USA 92, 4932–4936 (1995)
Hou, Y. M. & Schimmel, P. A simple structural feature is a major determinant of the identity of a transfer RNA. Nature 333, 140–145 (1988)
Ross, C. A. & Pickart, C. M. The ubiquitin–proteasome pathway in Parkinson's disease and other neurodegenerative diseases. Trends Cell Biol. 14, 703–711 (2004)
Welch, W. J. Role of quality control pathways in human diseases involving protein misfolding. Semin. Cell Dev. Biol. 15, 31–38 (2004)
Dobson, C. M. Principles of protein folding, misfolding and aggregation. Semin. Cell Dev. Biol. 15, 3–16 (2004)
Barral, J. M., Broadley, S. A., Schaffar, G. & Hartl, F. U. Roles of molecular chaperones in protein misfolding diseases. Semin. Cell Dev. Biol. 15, 17–29 (2004)
Muchowski, P. J. & Wacker, J. L. Modulation of neurodegeneration by molecular chaperones. Nature Rev. Neurosci. 6, 11–22 (2005)
Xu, C., Bailly-Maitre, B. & Reed, J. C. Endoplasmic reticulum stress: cell life and death decisions. J. Clin. Invest. 115, 2656–2664 (2005)
Zinszner, H. et al. CHOP is implicated in programmed cell death in response to impaired function of the endoplasmic reticulum. Genes Dev. 12, 982–995 (1998)
Murphy, R. M. Peptide aggregation in neurodegenerative disease. Annu. Rev. Biomed. Eng. 4, 155–174 (2002)
Sokabe, M., Okada, A., Yao, M., Nakashima, T. & Tanaka, I. Molecular basis of alanine discrimination in editing site. Proc. Natl Acad. Sci. USA 102, 11669–11674 (2005)
Ahel, I., Korencic, D., Ibba, M. & Soll, D. Trans-editing of mischarged tRNAs. Proc. Natl Acad. Sci. USA 100, 15422–15427 (2003)
Gatchel, J. R. & Zoghbi, H. Y. Diseases of unstable repeat expansion: mechanisms and common principles. Nature Rev. Genet. 6, 743–755 (2005)
Senderek, J. et al. Mutations in SIL1 cause Marinesco–Sjogren syndrome, a cerebellar ataxia with cataract and myopathy. Nature Genet. 37, 1312–1314 (2005)
Anttonen, A. K. et al. The gene disrupted in Marinesco–Sjogren syndrome encodes SIL1, an HSPA5 cochaperone. Nature Genet. 37, 1309–1311 (2005)
Zhao, L., Longo-Guess, C., Harris, B. S., Lee, J. W. & Ackerman, S. L. Protein accumulation and neurodegeneration in the woozy mutant mouse is caused by disruption of SIL1, a cochaperone of BiP. Nature Genet. 37, 974–979 (2005)
Ackerman, S. L. et al. The mouse rostral cerebellar malformation gene encodes an UNC-5-like protein. Nature 386, 838–842 (1997)
Hogan, B. Manipulating the Mouse Embryo: A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1986)
Xue, H., Shen, W. & Wong, J. T. Purification of hyperexpressed Bacillus subtilis tRNATrp cloned in Escherichia coli. J. Chromatogr. 613, 247–255 (1993)
Beebe, K., Merriman, E., Ribas De Pouplana, L. & Schimmel, P. A domain for editing by an archaebacterial tRNA synthetase. Proc. Natl Acad. Sci. USA 101, 5958–5963 (2004)
Pleiss, J. A. & Uhlenbeck, O. C. Identification of thermodynamically relevant interactions between EF-Tu and backbone elements of tRNA. J. Mol. Biol. 308, 895–905 (2001)
Swairjo, M. A. et al. Alanyl-tRNA synthetase crystal structure and design for acceptor-stem recognition. Mol. Cell 13, 829–841 (2004)
Nangle, L. A., Motta, C. M. & Schimmel, P. Global effects of mistranslation from an editing defect in mammalian cells. Chem. Biol. (in the press)
Acknowledgements
We thank The Jackson Laboratory sequencing, microchemistry, histology, bioimaging and microinjection services for their contributions. We also thank L. Dionne and K. Seburn for rotorod testing and data analysis, J. Szatkiewicz for assistance with statistical analysis, J. Torrance for graphics assistance, P. O'Maille, W. Waas and A. Wolfson for gifts of plasmids, and C. Motta and E. Merriman for facilitating the expression and purification of proteins. We are also grateful to the laboratory of J. Kelly for CD spectrometer assistance and to R. Burgess and P. Nishina for comments on the manuscript. This work was supported by grants from the National Institute of Neurological Disorders and Stroke and the National Institute on Aging to S.L.A., the National Institute of General Medical Sciences and a fellowship from the National Foundation for Cancer Research to P.S., a grant from the National Center for Research Resources to M.T.D., and an institutional National Cancer Institute core grant (JAX). S.L.A. is an investigator of the Howard Hughes Medical Institute. Author Contributions J.W.L. designed and performed mouse and cell culture experiments, K.B. and L.A.N. designed and performed biochemical analyses, J.J. performed mutation analysis, M.T.D. provided the congenic sti mice and oversaw initial mapping experiments, S.A.C. and C.M.L.-G. performed genetic mapping experiments, J.P.S. performed hair pathological analysis, P.S. and S.L.A. designed and supervised experiments. All authors discussed the results and commented on the manuscript, which was written by J.W.L., K.B., P.S. and S.L.A.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The sequence for mouse Aars has been deposited in GenBank under the accession number AY223875; sequence-tagged sites (STSs) for D8SlacCA1 (DQ386090), D8SlacCA2 (DQ386088) and D8SlacCA3 (DQ386089) can also be found in GenBank. Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.
Supplementary information
Supplemental Methods
This file contains additional details of the methods used in this study. (DOC 38 kb)
Supplementary Figure 1.
Pathway for acylated tRNAs entering the translational machinery. (PDF 838 kb)
Supplementary Figure 2.
Follicular dystrophy in sti/sti mutant mice. (PDF 3058 kb)
Supplementary Figure 3.
The rotorod latency to fall in sti/sti and wild type mice. (PDF 104 kb)
Supplementary Figure 4.
Genes in the sti critical region. (PDF 162 kb)
Supplementary Figure 5.
The sti mutation is in the alanyl-tRNA synthetase (Aars) gene. (PDF 862 kb)
Supplementary Figure 6.
Secondary structure analysis of mutant and wild type human AlaRS. (PDF 88 kb)
Supplementary Figure 7
Normal aminoacylation of tRNAAla by A734E AlaRS. (PDF 89 kb)
Supplementary Figure 8.
Normal deacylation of Ala-tRNAAla by A734E AlaRS (PDF 132 kb)
Supplementary Figure 9.
Inherent misacylation with serine and glycine by mouse AlaRS (PDF 172 kb)
Supplementary Figure 10.
Accumulation of misfolded proteins in sti/sti Purkinje cells. (PDF 1318 kb)
Rights and permissions
About this article
Cite this article
Lee, J., Beebe, K., Nangle, L. et al. Editing-defective tRNA synthetase causes protein misfolding and neurodegeneration. Nature 443, 50–55 (2006). https://doi.org/10.1038/nature05096
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nature05096
This article is cited by
-
Tryptophanyl-tRNA synthetase-1 (WARS-1) depletion and high tryptophan concentration lead to genomic instability in Caenorhabditis elegans
Cell Death Discovery (2024)
-
Phenylalanine-tRNA aminoacylation is compromised by ALS/FTD-associated C9orf72 C4G2 repeat RNA
Nature Communications (2023)
-
Geometric alignment of aminoacyl-tRNA relative to catalytic centers of the ribosome underpins accurate mRNA decoding
Nature Communications (2023)
-
The tRNA regulome in neurodevelopmental and neuropsychiatric disease
Molecular Psychiatry (2022)
-
Collisions and protein aggregations ahead: how aging affects ribosomal elongation dynamics
Signal Transduction and Targeted Therapy (2022)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.